Full Content is available to subscribers

Subscribe/Learn More  >
Proceedings Article

Membrane protein dynamics measured by two-photon ring correlation spectroscopy: theory and application to living cells

[+] Author Affiliations
Benedict Hebert, S. Elizabeth Hulme, Paul W. Wiseman

McGill Univ. (Canada)

Proc. SPIE 5700, Multiphoton Microscopy in the Biomedical Sciences V, 109 (April 11, 2005); doi:10.1117/12.593894
Text Size: A A A
From Conference Volume 5700

  • Multiphoton Microscopy in the Biomedical Sciences V
  • Ammasi Periasamy; Peter T. C. So
  • San Jose, CA | January 22, 2005

abstract

Many biochemical reactions and processes are regulated by proteins associated with cellular membranes. Trans-membrane proteins play an important role in many aspects of cellular development, cellular migration and signaling, and many diseases. Quantitative measurement of protein dynamics under various experimental conditions can give insights into the mechanisms of interaction and the functionality of the protein. Fluctuation techniques, such as fluorescence correlation spectroscopy (FCS) and image correlation spectroscopy (ICS), have been used for such dynamic measurements in membranes. However, FCS is limited to fast dynamics, and ICS works best on a flat 2-dimensional area. We present an alternative way to measure protein transport in spherical (non flat) living cells that combines laser scanning microscopy and image correlation methods: ring correlation spectroscopy (RCS). The RCS analysis is performed on CLSM or two-photon cross-sectional images of labeled proteins in the cell membrane, where the optical sectioning gives a "ring" of fluorescence in the images. We present computer simulations of two dimensional diffusion confined to the surface of a spherical shell, where the RCS analysis can extract the set input parameters from the simulation. As well, we present RCS analysis of two-photon microscopy images of Pre-B leukocytes cells expressing CD44 labeled with EGFP.

© (2005) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.
Citation

Benedict Hebert ; S. Elizabeth Hulme and Paul W. Wiseman
"Membrane protein dynamics measured by two-photon ring correlation spectroscopy: theory and application to living cells", Proc. SPIE 5700, Multiphoton Microscopy in the Biomedical Sciences V, 109 (April 11, 2005); doi:10.1117/12.593894; http://dx.doi.org/10.1117/12.593894


Access This Proceeding
Sign in or Create a personal account to Buy this proceeding ($15 for members, $18 for non-members).

Figures

Tables

NOTE:
Citing articles are presented as examples only. In non-demo SCM6 implementation, integration with CrossRef’s "Cited By" API will populate this tab (http://www.crossref.org/citedby.html).

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging & repositioning the boxes below.

Related Book Chapters

Topic Collections

Advertisement
  • Don't have an account?
  • Subscribe to the SPIE Digital Library
  • Create a FREE account to sign up for Digital Library content alerts and gain access to institutional subscriptions remotely.
Access This Proceeding
Sign in or Create a personal account to Buy this proceeding ($15 for members, $18 for non-members).
Access This Proceeding
Sign in or Create a personal account to Buy this article ($15 for members, $18 for non-members).
Access This Chapter

Access to SPIE eBooks is limited to subscribing institutions and is not available as part of a personal subscription. Print or electronic versions of individual SPIE books may be purchased via SPIE.org.